Nucleolin participates in DNA double-strand break-induced damage response through MDC1-dependent pathway.
Nucleolin participates in DNA double-strand break-induced damage response through MDC1-dependent pathway.
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DOI：
10.1371/journal.pone.0049245
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Komatsu K
中科院分区：
文献类型：
--
作者：
Kobayashi J;Fujimoto H;Sato J;Hayashi I;Burma S;Matsuura S;Chen DJ;Komatsu K
H2 AX是染色质重塑的重要因子，促进DNA损伤相关蛋白在DNA双链断裂（DSB）位点的积累。为了进一步了解H2 AX在DNA损伤反应（DDR）中的作用，我们试图通过蛋白质组学分析来鉴定H2 AX相互作用蛋白。因此，我们将核仁素确定为候选分子之一。在这里，我们展示了一个新的作用的主要核仁蛋白，核仁素，在DDR。核仁素与γ-H2 AX相互作用并聚集到激光微辐射DSB损伤位点。染色质免疫沉淀试验也显示核仁素在DSB位点周围聚集。核仁素缺失的细胞在暴露于γ射线后表现出对ATM依赖性磷酸化和随后的细胞周期检查点激活的抑制。此外，核仁素敲低降低HR和NHEJ活性，并显示IR诱导的HR/NHEJ因子的染色质积累减少，与γ-H2 AX焦点的延迟动力学一致。此外，核仁素敲低减少MDC 1相关事件，如53 BP 1，RNF 168，磷酸化ATM和H2 A泛素化的焦点形成。核仁素也表现出类似FACT-like活性的DSB损伤诱导的组蛋白驱逐染色质。总之，核仁素可以通过其FACT样功能在MDC 1相关通路中发挥作用，从而促进ATM依赖性细胞周期检查点和DSB修复。
H2AX is an important factor for chromatin remodeling to facilitate accumulation of DNA damage-related proteins at DNA double-strand break (DSB) sites. In order to further understand the role of H2AX in the DNA damage response (DDR), we attempted to identify H2AX-interacting proteins by proteomics analysis. As a result, we identified nucleolin as one of candidates. Here, we show a novel role of a major nucleolar protein, nucleolin, in DDR. Nucleolin interacted with γ-H2AX and accumulated to laser micro-irradiated DSB damage sites. Chromatin Immunoprecipitation assay also displayed the accumulation of nucleolin around DSB sites. Nucleolin-depleted cells exhibited repression of both ATM-dependent phosphorylation following exposure to γ-ray and subsequent cell cycle checkpoint activation. Furthermore, nucleolin-knockdown reduced HR and NHEJ activity and showed decrease in IR-induced chromatin accumulation of HR/NHEJ factors, agreeing with the delayed kinetics of γ-H2AX focus. Moreover, nucleolin-knockdown decreased MDC1-related events such as focus formation of 53 BP1, RNF168, phosphorylated ATM, and H2A ubiquitination. Nucleolin also showed FACT-like activity for DSB damage-induced histone eviction from chromatin. Taken together, nucleolin could promote both ATM-dependent cell cycle checkpoint and DSB repair by functioning in an MDC1-related pathway through its FACT-like function.
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